• Title/Summary/Keyword: Spore Production

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Studies on the Production of $\beta$-Galactosidase by Lactobacillus sporogenes - Production of Extracellular $\beta$- Galactosidase - (Lectobacillus sporogenes에 의한 $\beta$-Galactosidase 생산에 관한 연구 ( I ) -균체외 $\beta$-Galactosidase의 생산 -)

  • 김영만;이정치;정필근;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.59-66
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    • 1983
  • Cultural conditions for the production of extracellar $\beta$-galactosidase by Loctobacillus sporogenes, a spore forming lactic acid bacterium, were investigated with shaken flask and jar fermenter cultures. The fermentation medium giving maximum $\beta$-galactosidase yield was found to consist of 1 % lactose as a carbon source, 1.5% peptone as an organic nitrogen source. 0.2% ammonium sulfate as an inorganic nitrogen source, 0.8% ammonium phosphate dibasic as a phosphorus source, and 0.05% potassium chloride and 0.001% ferric chloride as mineral source. Optimal initial pH of the medium was 7.0 and the highest enzyme excretion was observed after 40 hours of cultivation at 37$^{\circ}C$. In this experiment, the 500$m\ell$ conical flask containing 50-200$m\ell$ of medium was shaken at 140 strokes per minute with 7cm amplitude in a reciprocating shaker. The maximum enzyme value attained was 38 U/$m\ell$ of the culture broth which was found to be slightly higher than the highest intermolecular enzyme activity (30 U/$m\ell$) observed after 24 hours of incubation. In the fermentor culture, the fermentation profile was shown to be similar to that observed in the shaken flask experiment. But the maximum extracellular enzyme activity was 45 U/$m\ell$ to be even higher than the value obtained with the shaken flask culture.

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Ultrastructure of the Cell Wall of a Null Pigmentation Mutant, npgA1, in Aspergillus nidulans

  • Chung, Yun-Shin;Kim, Jung-Mi;Han, Dong-Min;Chae, Keon-Sang;Jahng, Kwang-Yeop
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.224-231
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    • 2003
  • The null pigmentation mutant (npgA1) of Aspergillus nidulans was previously characterized by its production of no pigment at any stage of its life cycle, its reduction in hyphal branching, and its delay in the asexual spore development. The chemical composition of the cell wall was also altered in npgA1 mutants that became more sensitive to Novozyme 234$\^$TM/, which is possibly due to a structural defect in the cell wall. To investigate the effects of the cell wall structure on these pleiomorphic phenomena, we examined the ultrastructure of the cell wall in the npgA1 mutant (WX17). Scanning electron micrographs (SEM) showed that after being cultured for six days, the outermost layer of the conidial wall of WX17 peeled off. Although this phenotype suggested that the cell wall structure in WX17 may be modified, examination using TEM of the fine structure of cross-sectioned hyphal wall of WX17 did not show any differences from that of FGSC4. However, staining for carbohydrates of wall layers showed that the electron-translucent layer of the cell wall was missing in WX17. In addition, the outermost layer H1 of the hyphal wall was also absent in WX17. The ultrastructural observation and cytochemical analysis of cell walls suggested that the pigmentation defect in WX17 may be attributed to the lack of a layer in the cell wall.

Isolation and Identification of Pathogenic Microorganisms from Soybean Sprouts

  • Kim, Hye-Jung;Koo, Kyoung-Mo;Kim, Gi-Nahm;Lee, Dong-Sun;Paik, Hyun-Dong
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.305-309
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    • 2002
  • Raw soybean sprouts were tested for contamination with the following bacteria which have potential for pathogenesis or food spoilage : Salmonella spp., Escherichia coli O157:H7, Yersinia enterocolitica, Vibrio parahae-molyticus, Aeromonas hydrophila, Plesidomonas shigeloides, Pseudomonas aeruginosa, Staphylococcus aureus, Lis-teria monocytogenes, Bacillus cereus, Clostridium perfringens, Campylobacter jejuni, Erwinia spp., and Fusarium spp. Three of the above strains were isolated from the sprouts, and identified by morphological and biochemical methods including an API kit and ATB automated identification system. The isolate cultured in Cereus selective agar, a selective medium, was a Gram-positive, rod shaped, anaerobic spore former. The biochemical and culture tests revealed the following characteristics: catalase-positive, no growth on Simmon's citrate, NO₂ production and requirement of arginine for growth; the ATB automated identification system gave 99.8 % agreement for the identification of Bacillus cereus to the species level. The isolate cultured in Macconkey agar selective medium was Gram-negative, rod shaped and a gas former; the ATB-system gave 99.9% agreement for the identification of Aeromonas hydrophila to the species level. The isolate found in Pseudomonas isolation agar was Gram-negative, rod shaped, cytochrome oxidase-positive, a reducer of nitrates to nitrogen, and pyocyanin producer; the ATB-system gave 99.9 % agreement for the identification of Pseudomonas aeruginosa to the species level. These results indicate that the three bacteria species present in the soybean sprouts were Bacillus cereus, Aero-monas hydrophila, and Pseudomonas aeruginosa. Salmonella spp., Escherichia coli O157:H7, and Yersinia enter-ocolitica, which are associated with serious disease in humans, were not isolated from soybean sprouts examined in this study.

Production and Properties of Mannanase by a Bacillus amyloliquefaciens Isolate (Bacillus amyloliquefaciens 분리균의 Mannanase 생산성과 효소특성)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.50 no.2
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    • pp.158-163
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    • 2014
  • In the acidic LB plate, a bacterial strain was isolated from homemade soybean paste as a producer of the extracellular mannanase. The isolate YB-1402, which was a Gram-positive rod-shaped bacterium with spore, has been identified as Bacillus amyloliquefaciens on the basis of its 16S rDNA sequence and biochemical properties. Maximum mannanase productivity of the isolate YB-1402 was reached approximately 150 U/ml in LB broth supplemented with konjac (3.0%). The molecular mass of YB-1402 mannanase was estimated to approximately 38.0 kDa by zymogram of the culture filtrate on SDS-PAGE. The mannanase of culture filtrate was the most active at $55^{\circ}C$ and pH 5.5. The mannanase activity was completely maintained after pre-incubation at pH 3.0 to 10.0 for 1 h. The predominant products resulting from the mannanase hydrolysis were mannose, mannobiose and mannotriose for LBG or mannooligosaccharides. The enzyme could hydrolyze mannooligosaccharides larger than mannobiose.

Detection, Characterization and Antibiotic Susceptibility of Clostridioides (Clostridium) difficile in Meat Products

  • Muratoglu, Karlo;Akkaya, Esra;Hampikyan, Hamparsun;Bingol, Enver Baris;Cetin, Omer;Colak, Hilal
    • Food Science of Animal Resources
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    • v.40 no.4
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    • pp.578-587
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    • 2020
  • Clostridioides (Clostridium) difficile is a Gram (+), anaerobic, spore forming, rod shaped bacterium that can produce toxin. The objective of this study is to reveal the presence of C. difficile in meat products, to analyze the ribotype diversity by PCR and to evaluate the antibiotic susceptibility of isolated strains. The organism was isolated in 22 out of 319 (6.9%) examined meat product samples and 9 out of 22 (40.9%) isolates were identified as RT027 and all isolates had the ability of toxin production. In terms of antibiotic susceptibility, all isolates were susceptive to amoxicillin-clavulanic acid, tetracycline and vancomycin and 21 (95.4%) isolates to metronidazole. On the other hand, imipenem and cefotaxim resistance was observed in all. In conclusion, the results of this comprehensive study conducted in Turkey deduced the presence of C. difficile in different meat products. Therefore, these products can be evaluated as a potential contamination source of C. difficile from animals to humans especially for elders, youngsters, long terms wide spectrum antibiotic used and immuno-suppressed individuals.

Fermentation and Sporulation Characteristics of Saccharomyces cerevisiae SHY111 Isolated from Korean Traditional Rice Wine

  • Kim, Seung-Hwan;Chung, Oon-Chan;Woo, Im-Sun;Shin, Jae-Ho;Rho, Dong-Hyun;Rhee, In-Koo;Park, Heui-Dong
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.776-783
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    • 2000
  • Various alcohol yeast strains have been isolated from main mashes of Korean traditional liquors, and their genetic diversities were previously reported [23]. In this study, the strain SHY111, showing the highest alcohol production, was tested for its fermentation and sporulation characteristics. Additionally, its haploid cells were isolated and tested for their growth and fermentation patterns. The strain was identified as Saccharomyces cerevisiae based on its morphological and physiological characteristics. The sequences of the ITS(internal transcribed spacer) and 5.8S rDNA regions of S. cerevisiae SHY111 were found to be identical to those of S. cerevisiae that was obtained from through the yeast genome project. The maximum fermentation ratio obtained by the strain SHY111 (96.7%) was almost the same as that by S. cerevisiae Balyun No. 1 (96.5%) that was a little higher than that by S. cerevisiae KCCM11215(95.8%). The strain was induced for sporulation in a sporulation liquid medium using log phase cells grown in different types of pre-sporulation media, and its haploid cells were obtained by spore dissection using a micromanipulator. The majority of the spores formed a small colony on a YPD agar plate, and the haploid yeast cells derived from the strain SHY111 showed a variety of growth and alcohol fermentation patterns. It was proposed that the fermentation patterns were related to their growth phenotypes in the most haploid strains, but possible not in some strains.

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Cultural Characteristics of Mycelial Growth by Cordyceps militaris (번데기동충하초(Cordyceps militaris)의 균사 생장)

  • Sung, Jae-Mo;Choi, Young-Sang;Shrestha, Bhushan;Park, Young-Joon
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.1-5
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    • 2002
  • Cultural characteristics of Cordyceps militaris preserved in EFCC, Kangwon National University were investigated for the mass production. The higher mycelial density of C. militaris was observed in Sabouraud's yeast and Yeast Malt agars, but the higher mycelial growth in Mushroom Minimal agar than other agars. The mycelium of C. militaris was observed to grow well at $25^{\circ}C$ and pH 6.0 respectively. The dextrose was found the best suitable energy source among the carbohydrates used for its mycelial growth, while the fructose or lactose observed to be well for mycelial growth. Hemoglobin was observed to be the best among the protein sources used for mycelial growth, while tryptone found to be the best in the spore formation. Similarly, the mycelial growth was best in mineral salts of $KH_2PO_4$ or $K_2HPO_4$ and the optimum C/N ratio was 100 : 1.

SUPPRESSION OF HYDROGEN CONSUMING BACTERIA IN ANAEROBIC HYDROGEN FERMENTATION

  • Park, Woo-Shin;Jang, Nam-J.;Hyun, Seung-H.;Kim, In-S.
    • Environmental Engineering Research
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    • v.10 no.4
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    • pp.181-190
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    • 2005
  • Severe loss or hydrogen occurred in most anaerobic hydrogen fermentation reactors. Several selected methods were applied to suppress the consumption of hydrogen and increase the potential of production. As the first trial, pH shock was applied. The pH of reactor was dropped nearly to 3.0 by stopping alkalinity supply and on]y feeding glucose (5 g/L-d). As the pH was increase to $4.8{\pm}0.2,$ the degradation pathway was derived to solventogenesis resulting in disappearance of hydrogen in the headspace. In the aspect of bacterial community, methanogens weren't detected after 22 and 35 day, respectively. Even though, however, there was no methanogenic bacterium detected with fluorescence in-situ hybridization (FISH) method, hydrogen loss still occurred in the reactor showing a continuous increase of acetate when the pH was increased to $5.5{\pm}0.2$. This result was suggesting the possibility of the survival of spore fanning acetogenic bacteria enduring the severely acidic pH. As an alternative and additive method, nitrate was added in a batch experiment. It resulted in the increase of maximum hydrogen fraction from 29 (blank) to 61 % $(500\;mg\;NO_3/L)$. However, unfortunately, the loss of hydrogen occurred right after the depletion of nitrate by denitrification. In order to prevent the loss entangled with acetate formation, $CO_2$ scavenging in the headspace was applied to the hydrogen fermentation with heat-treated sludge since it was the primer of acetogenesis. As the $CO_2$ scavenging was applied, the maximum fraction of hydrogen was enhanced from 68 % to 87 %. And the loss of hydrogen could be protected effectively.

A Plant Growth-Promoting Pseudomonas fluorescens GL20: Mechanism for Disease Suppression, Outer Membrane Receptors for Ferric Siderophore, and Genetic Improvement for Increased Biocontrol Efficacy

  • LIM, HO SEONG;JUNG MOK LEE;SANG DAL KIM
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.249-257
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    • 2002
  • Pseudomonas fluorescens GL20 is a plant growth-promoting rhizobacterium that produces a large amount of hydroxamate siderophore under iron-limited conditions. The strain GL20 considerably inhibited the spore germination and hyphal growth of a plant pathogenic fungus, Fusarium solani, when iron was limited, significantly suppressed the root-rot disease on beans caused by F. solani, and enhanced the plant growth. The mechanism for the beneficial effect of strain GL20 on the disease suppression was due to the siderophore production, evidenced by mutant strains derived from the strain. Analysis of the outer membrane protein profile revealed that the growth of strain GL20 induced the synthesis of specific iron-regulated outer membrane proteins with molecular masses of 85- and 90 kDa as the high-affinity receptors for the ferric siderophore. In addition, a cross-feeding assay revealed the presence of multiple inducible receptors for heterologous siderophores in the strain. In order to induce increased efficacy and potential in biological control of plant disease, a siderophore-overproducing mutant, GL20-S207, was prepared by NTG mutagenesis. The mutant GL20-S207 produced nearly 2.3 times more siderophore than the parent strain. In pot trials of beans with F. solani, the mutant increased plant growth up to 1.5 times compared with that of the parent strain. These results suggest that the plant growth-promoting P. fluorescens GL20 and the genetically bred P. fluorescens GL20-S207 can play an important role in the biological control of soil-borne plant diseases in the rhizosphere.

Application of Arbuscular Mycorrhizal Fungi during the Acclimatization of Alpinia purpurata to Induce Tolerance to Meloidogyne arenaria

  • da Silva Campos, Maryluce Albuquerque;da Silva, Fabio Sergio Barbosa;Yano-Melo, Adriana Mayumi;de Melo, Natoniel Franklin;Maia, Leonor Costa
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.329-336
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    • 2017
  • An experiment was conducted to evaluate the tolerance of micropropagated and mycorrhized alpinia plants to the parasite Meloidogyne arenaria. The experimental design was completely randomized with a factorial arrangement of four inoculation treatments with arbuscular mycorrhizal fungi (AMF) (Gigaspora albida, Claroideoglomus etunicatum, Acaulospora longula, and a non-inoculated control) in the presence or absence of M. arenaria with five replicates. The following characteristics were evaluated after 270 days of mycorrhization and 170 days of M. arenaria inoculation: height, number of leaves and tillers, fresh mass of aerial and subterranean parts, dry mass of aerial parts, foliar area, nutritional content, mycorrhizal colonization, AMF sporulation, and the number of galls, egg masses, and eggs. The results indicated a significant interaction between the treatments for AMF spore density, total mycorrhizal colonization, and nutrient content (Zn, Na, and N), while the remaining parameters were influenced by either AMF or nematodes. Plants inoculated with A. longula or C. etunicatum exhibited greater growth than the control. Lower N content was observed in plants inoculated with AMF, while Zn and Na were found in larger quantities in plants inoculated with C. etunicatum. Fewer galls were observed on mycorrhized plants, and egg mass production and the number of eggs were lower in plants inoculated with G. albida. Plants inoculated with A. longula showed a higher percentage of total mycorrhizal colonization in the presence of the nematode. Therefore, the association of micropropagated alpinia plants and A. longula enhanced tolerance to parasitism by M. arenaria.